Od for preparation of cardiomyocyte hypoxia/ reoxygenation. When studying drug efficacy, in vitro experiments are often adopted to make highthroughput screening. On account of complex chemical composition and metabolic process of drug, basic use of monomer composition or extract to straight add into in vitro experiment system, it may influence the authenticity and reliability of experiment outcomes. The pharmacological technique of employing animal drugcontaining plasma to make in vitro experiments is always to give drug to animal for particular time and take its plasma to make in vitro experiments, which is closer to actual in vivo predicament, can be a sensible in vitro experiment method to study pharmacological efficacy, and may not only reflect the direct impact on the absorbable part in drug, but additionally can reflect the effects of the metabolites formed by drug composition in physique and drug induced endogenous active substances, closer to the genuine approach created by drug in physique atmosphere, and providing a brand new strategy for studying Chinese medicine pharmacology from cellular and molecular levels. LDH could be the sign enzyme of myocardial cells, When membrane structure of cardiac muscle adjust, integrity destruction or permeability improve, a large level of LDH of cells will leak, so the LDH content within the medium raise. A very good process which can react the harm price from the cell membrane indirectly ATPase is protease that exists in cell membrane andNote: 1 = the optimal concentration of ferulic acid, 2 = the optimal concentration of ferulic acid containing plasma, Compare to standard manage group, *P 0.later the digestion time may be selected at 7 eight min, till the tissue is absolutely digested and turned into translucent state. As well extended time and also higher temperature for digestion will make cardiomyocytes lose activity. pH is adjusted within the variety of 7.2 7.4, and higher or reduce will influence the regular growth of cardiomyocytes and weaken or lose the ability of spontaneous beating.1212086-74-2 Purity Too lengthy time or as well significant force of centrifugation will cause mechanical injury of cardiomyocytes, so 1000 r/min centrifugation for 10 min is acceptable. Soon after purification with all the differential adherent separation system, 5bromoa deoxyuridine is added to inhibit the synthesis of fibroblast DNA and protein, and by inhibiting the proliferation in the fibroblasts, it could make sure that cardiomyocytes still maintain higher purity after cultured for 3 five d. Since in vivo model operation is fairly complex, and modeled animal survival rate is reasonably low, so cells are generally chosen because the primary suggests for efficacy screening, and neonatal rat cardiomyocyte hypoxia/ reoxygenation is usually employed to simulate in vivo myocardial ischemia/reperfusion injury.874-20-4 Formula Xu Daqiang etc[24] and Wang Feng,[25] Chen Yupei etc[26] Firstly use 99.PMID:23849184 99 nitrogen to presaturate medium for 10 min (nitrogen 1 L/min), and then fill with 99.99 nitrogen once again for 30 s, drive away airPharmacognosy Magazine | July-September 2013 | Vol 9 | IssueRen, et al.: Protective effects of ferulic acid on major cultured neonatal rat cardiomyocytesorganelles membrane, its dynamic react the cell power metabolism plus the loss of function. The activity of Na+K+ATPase within the hypoxia/reoxygenation injury myocardial cells is decrease. Therefore that causes abnormalities of ion transport of inside and outside of cell, and tends to make the Na+ in cells unable to discharge, along with the gradient of Na+ across the plasma membrane descent, and activates Na+/Ca2.